Somatic cells can be used for rescuing wild mammals of ecological and economic importance, such as red-rumped agouti, through their application in advanced technologies. Thus, appropriate cell isolation, culture, and storage through cryopreservation can ensure the future safe use of these cells. We aimed to establish and evaluate the effects of culture time (second, fifth, and eighth passages) and cryopreservation on the morphology, viability, metabolism, proliferative activity, reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔΨm), and apoptosis on somatic cells derived from red-rumped agouti skin. Initially, we identified six dermal fibroblast lines by morphology, immunophenotyping, and karyotyping assays. In vitro culture after the second, fifth, and eighth passages, as well as the cryopreservation conditions used did not affect the metabolism or level of apoptosis. Nevertheless, cells in the fifth passage featured a reduction in proliferative activity and an increase in ROS levels when compared to second and eighth passage cells. Moreover, cryopreservation resulted in reduced ΔΨm when compared to non-cryopreserved cells. Additionally, cryopreserved cells showed a reduction in viability immediately after thawing; nevertheless, the viability of these cells was re-established after 11 days of in vitro culture and was similar to that of non-cryopreserved cells. In conclusion, we have shown that viable fibroblasts can be obtained from red-rumped agouti skin, featuring minimal changes after eight passages in in vitro culture systems. Additionally, adjustments to the cryopreservation protocol are necessary to reduce cellular oxidative stress caused by low temperatures.

Milkability is defined as the ability of an animal to give a regular, complete, and rapid milk secretion by the mammary gland in response to a proper milking technique. The aim of the present study was to investigate the relationship of milkability pattern with milk yield and somatic cell score in buffaloes. Milk yield and milkability can be observed through the milk flow profiles recorded by an electronic milkmeter (Lactocorder). A total of 2,288 milk flow curves of Mediterranean Italian buffaloes were used for one-way analysis of variance, and eight milk emission patterns were studied. The most represented milk flow curve was type 3 (27.32%), followed by type 6 (17.79%) characterized by a very long plateau phase. The less represented curve was type 1 (4.41%) characterized by long lag time and low peak flow rate. According to analysis of variance, milk yield ranged from 2.21 to 5.22 kg per milking for types 1 and 6, respectively, whereas the peak flow rate was minimum (0.50 kg/min) and maximum (1.73 kg/min) for types 1 and 4, respectively. The total milking time was on average 11.29 ± 3.68 min; lag time and milk emission time averaged 2.19 ± 2.34 min and 4.30 ± 2.33 min, respectively. The 12.5% (n = 286) of total curves were classified as bimodal and 60 of these were found in type 4. Based on literature, type 4 curves are representative of very short teat canals and very high milk flow. Average somatic cell score was 3.63 ± 1.67 units, with maximum least-squares mean found for type 1 and minimum for type 6. Buffaloes showing curves of type 5 and 6 were characterized by the greatest milk yield at milking, lowest somatic cell score, and lowest milking time. Results of the present study evidenced that such traits could be used in the dairy buffaloes as indicators to improve udder health and milkability.

The effect of nonylphenol (NP; either alone or in combination with ionizing radiation) on the induction of DNA strand breaks in mouse somatic cells has been examined. Male and female mice were repeatedly irradiated with X-rays (0.05 or 0.10 Gy), injected with NP (25 or 50mg/kg bw), or both (0.05 Gy+25 mg/kg bw NP or 0.10 Gy+50 mg/kg bw NP), for 2 weeks, 5 days/week. Liver, spleen, femora, lungs and kidneys were removed from each animal for the comet assay. NP-induced DNA damage differed, depending on organ and sex. In male mice, NP induced damage in all organs examined; in females, only the kidneys were affected. The effect of irradiation alone was similar in females and males. Combined exposure of males to 0.05 Gy+25 mg/kg bw NP significantly reduced the level of DNA strand breaks, compared to the controls and to 25mg/kg bw NP alone, in the majority of organs. The higher doses significantly increased damage to DNA in all organs examined. Combined exposure of females to low doses of both agents significantly enhanced damage to DNA in bone marrow lymphocytes and in cells of the liver and kidneys, compared to controls. At 0.10 Gy+50 mg/kg bw NP, DNA damage was increased in organs except liver and spleen. Although NP alone may not be mutagenic in female mice, its co-administration with irradiation may increase DNA damage in some organs. In contrast, in male mice, damage was reduced after combined irradiation-NP exposure, compared to NP alone.